Application of high performance liquid chromatography to the separation of microbiologically active compounds from heated sugar solutions and Lactarius necator mushrooms

Dissertation

Tapani Suortti

Research output: ThesisDissertationMonograph

1 Citation (Scopus)

Abstract

This publication is a summary of seven research reports dealing with the application of High Performance Liquid Chromatography (HPLC) to the isolation of microbiologically active compounds in heated neutral glucose and fructose solutions and in aqueous extracts of the mushroom Lactarius necator.In both cases the starting point was a micro biologically measurable activity which was used as detection method during the isolation procedures.The formation of antimicrobial activity in neutral aqueous glucose and fructose solutions was the first object of study.Starting from the original solutions and by chromatographing them twice the compounds responsible for the antimicrobial activity could be recovered in crystalline form.The first separation step was an ordinary reverse phase chromatography, whereas the second was specific for these compounds and involved the application of a gel permeation chromatography column for reverse phase separation.As a result the compounds responsible for the antimicrobial activity were identified as various phenolic compounds.The aqueous extract of the edible mushroom Lactarius necator has been shown to exhibit strong mutagenic activity in the Ames Salmonella assay. The compound respon sible for this activity was isolated by HPLC This compound, designated here as necatorin was shown to be one of the strongest mutagens hitherto found in nature (6000 revertants/ g with the tester strain TA100).Necatorin was found to be rather stable, so that the normal methods applied in the preparation of mushrooms for consumption blanching, brining, pickling or freezing do not remove or inactivate it efficiently.Necatorin is a normal constituent of the mushroom with a rather high initial concentration (3...20 mg/kg fresh weight), and not an accidental contamination.In both the systems studied the application of HPLC in reverse phase mode made possible the use of the original microbial activity test as detector with minimal manipulation of the isolated fractions.Therefore investigations could be directed to the correct fractions in these complex mixtures.
Original languageEnglish
QualificationDoctor Degree
Awarding Institution
  • Helsinki University of Technology
Award date3 Jun 1986
Place of PublicationEspoo
Publisher
Print ISBNs951-38-2589-2
Publication statusPublished - 1986
MoE publication typeG4 Doctoral dissertation (monograph)

Fingerprint

High performance liquid chromatography
Sugars
Fructose
Brining
Blanching
Glucose
Pickling
Salmonella
Mutagens
Gel permeation chromatography
Chromatography
Complex Mixtures
Freezing
Phase separation
Assays
Contamination
Crystalline materials
Detectors
necatorin

Keywords

  • high performance
  • liquid chromatography
  • HPLC
  • extracts
  • Lactarius necator
  • mutagens
  • antiinfective agents

Cite this

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title = "Application of high performance liquid chromatography to the separation of microbiologically active compounds from heated sugar solutions and Lactarius necator mushrooms: Dissertation",
abstract = "This publication is a summary of seven research reports dealing with the application of High Performance Liquid Chromatography (HPLC) to the isolation of microbiologically active compounds in heated neutral glucose and fructose solutions and in aqueous extracts of the mushroom Lactarius necator.In both cases the starting point was a micro biologically measurable activity which was used as detection method during the isolation procedures.The formation of antimicrobial activity in neutral aqueous glucose and fructose solutions was the first object of study.Starting from the original solutions and by chromatographing them twice the compounds responsible for the antimicrobial activity could be recovered in crystalline form.The first separation step was an ordinary reverse phase chromatography, whereas the second was specific for these compounds and involved the application of a gel permeation chromatography column for reverse phase separation.As a result the compounds responsible for the antimicrobial activity were identified as various phenolic compounds.The aqueous extract of the edible mushroom Lactarius necator has been shown to exhibit strong mutagenic activity in the Ames Salmonella assay. The compound respon sible for this activity was isolated by HPLC This compound, designated here as necatorin was shown to be one of the strongest mutagens hitherto found in nature (6000 revertants/ g with the tester strain TA100).Necatorin was found to be rather stable, so that the normal methods applied in the preparation of mushrooms for consumption blanching, brining, pickling or freezing do not remove or inactivate it efficiently.Necatorin is a normal constituent of the mushroom with a rather high initial concentration (3...20 mg/kg fresh weight), and not an accidental contamination.In both the systems studied the application of HPLC in reverse phase mode made possible the use of the original microbial activity test as detector with minimal manipulation of the isolated fractions.Therefore investigations could be directed to the correct fractions in these complex mixtures.",
keywords = "high performance, liquid chromatography, HPLC, extracts, Lactarius necator, mutagens, antiinfective agents",
author = "Tapani Suortti",
year = "1986",
language = "English",
isbn = "951-38-2589-2",
series = "Publications / Technical Research Centre of Finland",
publisher = "VTT Technical Research Centre of Finland",
number = "28",
address = "Finland",
school = "Helsinki University of Technology",

}

Application of high performance liquid chromatography to the separation of microbiologically active compounds from heated sugar solutions and Lactarius necator mushrooms : Dissertation. / Suortti, Tapani.

Espoo : VTT Technical Research Centre of Finland, 1986. 65 p.

Research output: ThesisDissertationMonograph

TY - THES

T1 - Application of high performance liquid chromatography to the separation of microbiologically active compounds from heated sugar solutions and Lactarius necator mushrooms

T2 - Dissertation

AU - Suortti, Tapani

PY - 1986

Y1 - 1986

N2 - This publication is a summary of seven research reports dealing with the application of High Performance Liquid Chromatography (HPLC) to the isolation of microbiologically active compounds in heated neutral glucose and fructose solutions and in aqueous extracts of the mushroom Lactarius necator.In both cases the starting point was a micro biologically measurable activity which was used as detection method during the isolation procedures.The formation of antimicrobial activity in neutral aqueous glucose and fructose solutions was the first object of study.Starting from the original solutions and by chromatographing them twice the compounds responsible for the antimicrobial activity could be recovered in crystalline form.The first separation step was an ordinary reverse phase chromatography, whereas the second was specific for these compounds and involved the application of a gel permeation chromatography column for reverse phase separation.As a result the compounds responsible for the antimicrobial activity were identified as various phenolic compounds.The aqueous extract of the edible mushroom Lactarius necator has been shown to exhibit strong mutagenic activity in the Ames Salmonella assay. The compound respon sible for this activity was isolated by HPLC This compound, designated here as necatorin was shown to be one of the strongest mutagens hitherto found in nature (6000 revertants/ g with the tester strain TA100).Necatorin was found to be rather stable, so that the normal methods applied in the preparation of mushrooms for consumption blanching, brining, pickling or freezing do not remove or inactivate it efficiently.Necatorin is a normal constituent of the mushroom with a rather high initial concentration (3...20 mg/kg fresh weight), and not an accidental contamination.In both the systems studied the application of HPLC in reverse phase mode made possible the use of the original microbial activity test as detector with minimal manipulation of the isolated fractions.Therefore investigations could be directed to the correct fractions in these complex mixtures.

AB - This publication is a summary of seven research reports dealing with the application of High Performance Liquid Chromatography (HPLC) to the isolation of microbiologically active compounds in heated neutral glucose and fructose solutions and in aqueous extracts of the mushroom Lactarius necator.In both cases the starting point was a micro biologically measurable activity which was used as detection method during the isolation procedures.The formation of antimicrobial activity in neutral aqueous glucose and fructose solutions was the first object of study.Starting from the original solutions and by chromatographing them twice the compounds responsible for the antimicrobial activity could be recovered in crystalline form.The first separation step was an ordinary reverse phase chromatography, whereas the second was specific for these compounds and involved the application of a gel permeation chromatography column for reverse phase separation.As a result the compounds responsible for the antimicrobial activity were identified as various phenolic compounds.The aqueous extract of the edible mushroom Lactarius necator has been shown to exhibit strong mutagenic activity in the Ames Salmonella assay. The compound respon sible for this activity was isolated by HPLC This compound, designated here as necatorin was shown to be one of the strongest mutagens hitherto found in nature (6000 revertants/ g with the tester strain TA100).Necatorin was found to be rather stable, so that the normal methods applied in the preparation of mushrooms for consumption blanching, brining, pickling or freezing do not remove or inactivate it efficiently.Necatorin is a normal constituent of the mushroom with a rather high initial concentration (3...20 mg/kg fresh weight), and not an accidental contamination.In both the systems studied the application of HPLC in reverse phase mode made possible the use of the original microbial activity test as detector with minimal manipulation of the isolated fractions.Therefore investigations could be directed to the correct fractions in these complex mixtures.

KW - high performance

KW - liquid chromatography

KW - HPLC

KW - extracts

KW - Lactarius necator

KW - mutagens

KW - antiinfective agents

M3 - Dissertation

SN - 951-38-2589-2

T3 - Publications / Technical Research Centre of Finland

PB - VTT Technical Research Centre of Finland

CY - Espoo

ER -